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Sains Malaysiana 54(9)(2025): 2137-2149
http://doi.org/10.17576/jsm-2025-5409-03
Pengekstrakan dan Pengenalpastian Peptida Perencat α-Amilase daripada Benih Elateriospermum tapos
(Extraction and Identification of
α-Amylase Inhibitory Peptides from
PEI-GEE YAP, NAUFAL ARSHAD
& CHEE-YUEN GAN*
Pusat Penyelidikan Biokimia Analisis (ABrC), Kampus SAINS@USM, Universiti Sains Malaysia, Lebuh Bukit Jambul, 11900 Bayan
Lepas, Pulau Pinang, Malaysia
Received: 14 March 2025/Accepted: 16 July 2025
Abstrak
Elateriospermum tapos adalah tumbuhan unik yang berasal dari Asia Tenggara, digunakan secara tradisi untuk merawat penyakit kronik termasuk kencing manis. Uji kaji ini bertujuan untuk mengekstrak dan mengenal pasti peptida perencat α-amilase daripada benih E. tapos. Protein daripada benih E. tapos (ETSPI) telah dihidrolisis dengan menggunakan enzim Neutrase dan darjah hidrolisis tertinggi (34.09±2.50%) diperoleh dari nisbah enzim-ke-substrat 1:10 (b/b) selama 4 jam. Aktiviti perencatan α-amilase tertinggi (41.74±1.45%) diperoleh pada nisbah yang sama tetapi pada masa hidrolisis selama 2 jam. Ultrafiltrasi kemudian digunakan untuk memecahkan hidrolisat dengan pecahan <3 kDa memberikan potensi yang paling tinggi dalam perencatan α-amilase. Dalam pecahan ini, empat peptida: ME, LLAH, LLHA dan LLPL telah dikenal pasti dan dipilih melalui teknik bantuan enzim yang digabungkan dengan pendekatan spektrometri jisim. Peptida ini diramalkan sebagai tidak toksik dan bukan agregat melalui pendekatan bioinformatik. Kajian hubungan aktiviti struktur menunjukkan bahawa semua peptida terpilih menunjukkan interaksi yang signifikan (nilai p PepSite2<0.25) dengan α-amilase pankreas and
air liur manusia. Aktiviti perencatan α-amilase peptida telah disahkan dengan nilai IC50 adalah seperti berikut: LLAH (2.88±0.01 mg/mL), LLHA (8.05±0.07 mg/mL),
LLPL (4.22±0.01 mg/mL) dan ME (1.41±0.00 mg/mL). Penemuan ini menunjukkan kejayaan pengenalpastian peptida perencat α-amilase dengan potensi aplikasi dalam penemuan ubat dan industri nutraseutikal.
Kata kunci: α-amilase; anti-kencing manis; bioinformatik; peptida bioaktif; terapi semula jadi
Abstract
Elateriospermum tapos is a unique plant native
to Southeast Asia, traditionally used to treat chronic diseases including
diabetes mellitus. This experiment aimed to extract and identify α-amylase
inhibitor peptides from E. tapos seeds. E. tapos seed protein isolate (ETSPI) was hydrolysed using Neutrase enzyme, obtaining the highest degree of
hydrolysis (34.09±2.50%) at enzyme-to-substrate ratio of 1:10 (w/w) over 4 h.
The highest α-amylase inhibitory activity (41.74±1.45%) was obtained at
the same ratio but at a hydrolysis time of 2 h. Ultrafiltration was used to
fractionate the hydrolysate, in which the fraction <3 kDa gave the most potent potential in inhibiting α-amylase. In this fraction,
four peptides: ME, LLAH, LLHA, and LLPL were identified and selected through
the enzyme-assisted technique incorporated with tandem mass spectrometry
approach. These peptides were predicted as non-toxic and non-aggregate through
bioinformatics approach. Structure activity relationship study demonstrated
that all selected peptides showed significant (PepSite2 p-value<0.25)
interaction with human pancreatic and salivary α-amylases. The
α-amylase inhibitory activity of the peptides was validated whereby the IC50 values were as follows: LLAH (2.88±0.01 mg/mL), LLHA (8.05±0.07 mg/mL), LLPL
(4.22±0.01 mg/mL), and ME (1.41±0.00 mg/mL). These findings demonstrated
successful identification of α-amylase inhibitor peptides with potential
applications in drug discovery and the nutraceutical industry.
Keywords: α-amylase; anti-diabetes;
bioactive peptide; bioinformatics; natural therapeutics
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